Characterization of nerve growth factors (NGFs) from snake venoms by use of a novel, quantitative bioassay utilizing pheochromocytoma (PC12) cells overexpressing human trkA receptors

Katzir, Itzhak; Shani, J.; Goshen, Gal; Sela, J.; Ninary, Einat; Dogonovski, Anne Marie; Shabashov, Dalia; Inoue, Shintaro; Ikeda, Kiyoshi; Hayashi, Kyozo; Gorinstein, Shela; Deutsch, Joseph; Lazarovici, Philip

doi:10.1016/s0041-0101(03)00225-3

Cited by 21 publications

(16 citation statements)

References 31 publications

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“…By contrast, viper NGF stimulated the migration of HAECs with a potency and an efficacy similar to that of rhVEGF. The higher potency of vNGF by comparison to the other NGFs may be attributed either to the Asn-21 glycosylation found only in vNGF (Katzir et al, 2003) but not in mouse and human recombinant NGF (Hayashi et al, 1996) or to an increased affinity toward TrkA receptors due to primary sequence changes compared with the other NGFs. It is well known that glycosylation of NGF and FGF results in an increased stability, probably causing more efficient and/or prolonged receptor stimulation (Delli-Bovi et al, 1988;Murphy et al, 1989).…”

Section: Discussionmentioning

confidence: 92%

“…Interestingly, the order of potency by which the various NGF analogs stimulated HAEC migration (vNGF Ͼ Ͼ mNGF Ͼ rhNGF) is opposite to the order by which these agonists induce neurite outgrowth (rhNGF Ͼ mNGF Ͼ vNGF) in PC12 cells overexpressing neuronal human recombinant TrkA receptors (Katzir et al, 2003). This observation suggests that the TrkA receptor in HAECs may be of another subtype than human neuronal TrkA.…”

Section: Discussionmentioning

confidence: 95%

“…Human recombinant nerve growth factor (rhNGF) and mouse ␤-nerve growth factor (2.5S-mNGF) were kindly supplied by Alomone Labs (Jerusalem, Israel). vNGF was purified as described previously (Hayashi et al, 1996;Katzir et al, 2003). Stock solutions of the different NGFs were routinely analyzed for activity in the PC12 bioassay (Katzir et al, 2003).…”

Section: Methodsmentioning

confidence: 99%

“…vNGF was purified as described previously (Hayashi et al, 1996;Katzir et al, 2003). Stock solutions of the different NGFs were routinely analyzed for activity in the PC12 bioassay (Katzir et al, 2003). Stock solutions of all growth factors (0.2-2.2 mg/ml) in PBS were aliquoted and stored at Ϫ20°C.…”

Section: Methodsmentioning

confidence: 99%

“…The majority of research on NGF has been performed using NGF isolated from the male mouse submaxillary gland. More recently, human recombinant NGF (Rask, 1999) and snake venom NGFs (Hayashi et al, 1996;Katzir et al, 2003) have become important additional NGF agonist tools. Several recent reports indicate that NGF exerts a variety of effects on peripheral tissues, including the vasculature, suggesting that NGF may be a novel angiogenic factor (Cantarella et al, 2002;Lazarovici et al, 2005).…”

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confidence: 99%

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Nerve Growth Factor-Induced Migration of Endothelial Cells

Dollé¹,

Rezvan²,

Allen³

et al. 2005

The Journal of Pharmacology and Experimental Therapeutics

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Nerve growth factor (NGF) is a well known neurotropic and neurotrophic agonist in the nervous system, which recently was shown to also induce angiogenic effects in endothelial cells (ECs). To measure NGF effects on the migration of cultured ECs, an important step in neoangiogenesis, we optimized an omnidirectional migration assay using human aortic endothelial cells (HAECs) and validated the assay with human recombinant basic fibroblast growth factor (rhbFGF) and human recombinant vascular endothelial growth factor (rhVEGF). The potencies of nerve growth factor purified from various species (viper, mouse, and recombinant human) to stimulate HAEC migration was similar to that of VEGF and basic fibroblast growth factor (bFGF) (EC 50 of ϳ0.5 ng/ml). Recombinant human bFGF was significantly more efficacious than either viper NGF or rhVEGF, both of which stimulated HAEC migration by ϳ30% over basal spontaneous migration. NGF-mediated stimulation of HAEC migration was completely blocked by the NGF/TrkA receptor antagonist K252a [(8R*,9S*,11S*)-(ı)-9-hydroxy-9-methoxycarbonyl-8-methyl-2,3,9,10-tetrahydro-8,11-epoxy-1H,-8H,11H-2,7b,11a-triazadibenzo(a,g)cycloocta(c,d,e)trindene-1-one] (30 nM) but not by the VEGF/Flk receptor antagonist ) methylidenyl]-indolin-2-one] (250 nM), indicating a direct effect of NGF via TrkA receptor activation on HAEC migration. Viper NGF stimulation of HAEC migration was additively increased by either rhVEGF or rhbFGF, suggesting a potentiating interaction between their tyrosine kinase receptor signaling pathways. Viper NGF represents a novel pharmacological tool to investigate possible TrkA receptor subtypes in endothelial cells. The ability of NGF to stimulate migration of HAEC cells in vitro implies that this factor may play an important role in the cardiovascular system besides its well known effects in the nervous system.

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Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 95%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Nerve Growth Factor-Induced Migration of Endothelial Cells

Dollé¹,

Rezvan²,

Allen³

et al. 2005

The Journal of Pharmacology and Experimental Therapeutics

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Post-translational modification accounts for the presence of varied forms of nerve growth factor in Australian elapid snake venoms

et al. 2006

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The Australian elapid snakes are amongst the most venomous snakes in the world, but much less is known about the overall venom composition in comparison to Asian and American snakes. We have used a combined approach of cDNA cloning and 2-DE with MS to identify nerve growth factor (NGF) in venoms of the Australian elapid snakes and demonstrate its neurite outgrowth activity. While a single 730 nucleotide ORF, coding for a 243 amino acid precursor protein was detected in all snakes, use of 2-DE identified NGF proteins with considerable variation in molecular size within and between the different snakes. The variation in size can be explained at least in part by N-linked glycosylation. It is possible that these modifications alter the stability, activity and other characteristics of the snake NGFs. Further characterisation is necessary to delineate the function of the individual NGF isoforms.

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Urea-Induced Dissociation of Nerve Growth Factor from Venom of Chinese Cobra by SEC

Bian

2014

Chromatographia

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Characterization of nerve growth factors (NGFs) from snake venoms by use of a novel, quantitative bioassay utilizing pheochromocytoma (PC12) cells overexpressing human trkA receptors

Cited by 21 publications

References 31 publications

Nerve Growth Factor-Induced Migration of Endothelial Cells

Nerve Growth Factor-Induced Migration of Endothelial Cells

Post-translational modification accounts for the presence of varied forms of nerve growth factor in Australian elapid snake venoms

Urea-Induced Dissociation of Nerve Growth Factor from Venom of Chinese Cobra by SEC

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